21, 24 In this study, administration of saffron to DEN-treated ra

21, 24 In this study, administration of saffron to DEN-treated rats reversed DEN-induced up-regulation of NF-κB-p65 subunit (Fig. 4). A similar result is reported in our in vitro studies, where saffron treatment caused an early decrease in the phosphorylation state of IκB (Fig. 5D). This anti-inflammatory effect of saffron against acute and chronic models of inflammation has been previously reported as well.8

In summary, the data presented here show that saffron dramatically inhibited both nodular and FAH formation in livers of DEN-treated rats. This inhibition was associated with induced apoptosis, reduced cell proliferation, decreased oxidative stress and down-regulation of inflammatory markers, such as COX-2, iNOS, NF-κB-p65 and p-TNFR1 INCB024360 nmr expressions. Figure 6 incorporates our data into a model showing a possible mechanism of the anticancer protective effect of saffron by promoting apoptosis, inhibiting cell proliferation, and blocking inflammation in hepatocarcinomas. Further investigations are currently underway to investigate in more detail the mechanism of action of saffron extract. This work was financially supported by Emirates Foundation grant 2009-079 to A.A. Authors are grateful to Aktham Awwad (Tawam Hospital), Rkia Al-Kharrge (UAE University) for assessing hepatic nodules. Authors are also indebted to Hamdi Kandil (UAE University) and Moustafa Abdalla (Groves High School, MI USA) for their technical assistance.

Additional selleck chemical Supporting Information may be found in the online version of this article. “
“See article in J. Gastroenterol. Hepatol. 2010; 25: 1087–1092 External pancreatic fistula (EPF), also called pancreaticocutaneous fistula, is a reported complication in 38% of patients undergoing pancreatic resection,1 10% after necrosectomy, and 20% after pseudocyst drainage.2 Thus, EPF results MCE公司 from injury during pancreatic resection for pancreaticoduodenal trauma, debridement for necrotizing pancreatitis, and percutaneous drainage for communicating pseudocysts. Although the exact pathogenesis is unclear, parenchymal necrosis that might disrupt the small or even the larger pancreatic ducts is considered as

the most crucial factor.3,4 Clinically, most patients with an EPF present with fistulas pouring low or moderate output volumes that are not life threatening,3 but occasionally serious complications occur. These include abscess, bleeding from the fistulous tract, and sepsis secondary to abscess formation; the latter is usually associated with high fistulous output, and mortality in such complicated cases is 13%–36%.3–5 In patients afflicted with low fistulous output EPF, conservative management is appropriate. This includes nil by mouth, total parenteral nutrition, and administration of inhibitors of pancreatic secretion, such as somatostatin or its analog. Such ‘conservative management’ leads to spontaneous closure of the fistula in 40%–90% of cases.

21, 24 In this study, administration of saffron to DEN-treated ra

21, 24 In this study, administration of saffron to DEN-treated rats reversed DEN-induced up-regulation of NF-κB-p65 subunit (Fig. 4). A similar result is reported in our in vitro studies, where saffron treatment caused an early decrease in the phosphorylation state of IκB (Fig. 5D). This anti-inflammatory effect of saffron against acute and chronic models of inflammation has been previously reported as well.8

In summary, the data presented here show that saffron dramatically inhibited both nodular and FAH formation in livers of DEN-treated rats. This inhibition was associated with induced apoptosis, reduced cell proliferation, decreased oxidative stress and down-regulation of inflammatory markers, such as COX-2, iNOS, NF-κB-p65 and p-TNFR1 find more expressions. Figure 6 incorporates our data into a model showing a possible mechanism of the anticancer protective effect of saffron by promoting apoptosis, inhibiting cell proliferation, and blocking inflammation in hepatocarcinomas. Further investigations are currently underway to investigate in more detail the mechanism of action of saffron extract. This work was financially supported by Emirates Foundation grant 2009-079 to A.A. Authors are grateful to Aktham Awwad (Tawam Hospital), Rkia Al-Kharrge (UAE University) for assessing hepatic nodules. Authors are also indebted to Hamdi Kandil (UAE University) and Moustafa Abdalla (Groves High School, MI USA) for their technical assistance.

Additional see more Supporting Information may be found in the online version of this article. “
“See article in J. Gastroenterol. Hepatol. 2010; 25: 1087–1092 External pancreatic fistula (EPF), also called pancreaticocutaneous fistula, is a reported complication in 38% of patients undergoing pancreatic resection,1 10% after necrosectomy, and 20% after pseudocyst drainage.2 Thus, EPF results MCE from injury during pancreatic resection for pancreaticoduodenal trauma, debridement for necrotizing pancreatitis, and percutaneous drainage for communicating pseudocysts. Although the exact pathogenesis is unclear, parenchymal necrosis that might disrupt the small or even the larger pancreatic ducts is considered as

the most crucial factor.3,4 Clinically, most patients with an EPF present with fistulas pouring low or moderate output volumes that are not life threatening,3 but occasionally serious complications occur. These include abscess, bleeding from the fistulous tract, and sepsis secondary to abscess formation; the latter is usually associated with high fistulous output, and mortality in such complicated cases is 13%–36%.3–5 In patients afflicted with low fistulous output EPF, conservative management is appropriate. This includes nil by mouth, total parenteral nutrition, and administration of inhibitors of pancreatic secretion, such as somatostatin or its analog. Such ‘conservative management’ leads to spontaneous closure of the fistula in 40%–90% of cases.

21, 24 In this study, administration of saffron to DEN-treated ra

21, 24 In this study, administration of saffron to DEN-treated rats reversed DEN-induced up-regulation of NF-κB-p65 subunit (Fig. 4). A similar result is reported in our in vitro studies, where saffron treatment caused an early decrease in the phosphorylation state of IκB (Fig. 5D). This anti-inflammatory effect of saffron against acute and chronic models of inflammation has been previously reported as well.8

In summary, the data presented here show that saffron dramatically inhibited both nodular and FAH formation in livers of DEN-treated rats. This inhibition was associated with induced apoptosis, reduced cell proliferation, decreased oxidative stress and down-regulation of inflammatory markers, such as COX-2, iNOS, NF-κB-p65 and p-TNFR1 AZD2014 in vivo expressions. Figure 6 incorporates our data into a model showing a possible mechanism of the anticancer protective effect of saffron by promoting apoptosis, inhibiting cell proliferation, and blocking inflammation in hepatocarcinomas. Further investigations are currently underway to investigate in more detail the mechanism of action of saffron extract. This work was financially supported by Emirates Foundation grant 2009-079 to A.A. Authors are grateful to Aktham Awwad (Tawam Hospital), Rkia Al-Kharrge (UAE University) for assessing hepatic nodules. Authors are also indebted to Hamdi Kandil (UAE University) and Moustafa Abdalla (Groves High School, MI USA) for their technical assistance.

Additional Selleckchem Carfilzomib Supporting Information may be found in the online version of this article. “
“See article in J. Gastroenterol. Hepatol. 2010; 25: 1087–1092 External pancreatic fistula (EPF), also called pancreaticocutaneous fistula, is a reported complication in 38% of patients undergoing pancreatic resection,1 10% after necrosectomy, and 20% after pseudocyst drainage.2 Thus, EPF results medchemexpress from injury during pancreatic resection for pancreaticoduodenal trauma, debridement for necrotizing pancreatitis, and percutaneous drainage for communicating pseudocysts. Although the exact pathogenesis is unclear, parenchymal necrosis that might disrupt the small or even the larger pancreatic ducts is considered as

the most crucial factor.3,4 Clinically, most patients with an EPF present with fistulas pouring low or moderate output volumes that are not life threatening,3 but occasionally serious complications occur. These include abscess, bleeding from the fistulous tract, and sepsis secondary to abscess formation; the latter is usually associated with high fistulous output, and mortality in such complicated cases is 13%–36%.3–5 In patients afflicted with low fistulous output EPF, conservative management is appropriate. This includes nil by mouth, total parenteral nutrition, and administration of inhibitors of pancreatic secretion, such as somatostatin or its analog. Such ‘conservative management’ leads to spontaneous closure of the fistula in 40%–90% of cases.

For each of these three comparisons, the difference in responder

For each of these three comparisons, the difference in responder rate and associated 95% confidence interval was determined. Once the optimum TBV dose was identified, a test of noninferior efficacy was performed by comparing the proportions of responders at TW12 in the MLN0128 ic50 optimal TBV and RBV treatment arms. Chi-squared or the Fisher’s exact test compared anemia rates between the TBV and RBV groups with a 95% confidence interval. Secondary efficacy measures included the SVR defined as HCV RNA <100 copies/mL (39 IU/mL) and/or at least a 2-log decrease from baseline at TW4, TW24, TW48 and FW4 and FW12 and relapse rates at FW4, FW12, and

FW24. Secondary safety measures included the comparison of incidence of treatment-emergent adverse events. Subgroup analysis by HCV RNA levels at baseline, body weight, age, sex, race, and baseline fibrosis were performed using the trend test and the Fisher’s exact test for the primary endpoint. In addition, the Cochran-Mantel-Haenszel check details procedure, with the

Breslow-Day test was used to examine the homogeneity of treatment effect across strata. The investigators and the sponsor managed the data for this study. The sponsor completed the statistical analysis. The authors had access to the clinical study report and have either written or provided intellectual input to the manuscript. A total of 278 patients were randomized at 51 U.S. centers between March 2007 and October 2008. A total of 86 (41%) of patients in the TBV arms and 25 (36%) in the RBV arm completed treatment and follow-up. Overall, 122 (59%) patients withdrew prematurely

in the TBV arms compared to 45 (64%) in the RBV group. The most commonly cited reasons for premature withdrawal were lack of response (29%) and adverse events (20%). Figure 1 shows the disposition of patients during treatment. Baseline characteristics across the four treatment groups were similar (Table 1). The majority of patients were male (61%) with a mean weight of 82.1 kg and mean age of 49 years. African American or Latino patients accounted for 30% of the study medchemexpress population and 81% had high viral load defined as >400,000 IU/mL at baseline. The proportions of patients in the ITT population with an EVR, the primary endpoint of this study, were comparable between all groups with no statistical difference versus RBV. EVR was achieved in 64.2% (43 of 67) in the 20 mg/kg group, 57.1% (40 of 70) in the 25 mg/kg group, 54.4% (37 of 68) of the 30 mg/kg group and 51.4% (36 of 70) in the RBV group. Virologic response for TW4, 12, 24, and 48 as well as SVR are shown in Table 2. The proportion of patients with undetectable HCV RNA at every time point was similar between the TBV and RBV groups. Although responder rates were numerically lower at TW12 in the TBV 30 mg/kg group and somewhat higher at TW24 and TW48 in the TBV 20 mg/kg group, they were not significantly different for any of the TBV doses compared with RBV.

For each of these three comparisons, the difference in responder

For each of these three comparisons, the difference in responder rate and associated 95% confidence interval was determined. Once the optimum TBV dose was identified, a test of noninferior efficacy was performed by comparing the proportions of responders at TW12 in the Deforolimus clinical trial optimal TBV and RBV treatment arms. Chi-squared or the Fisher’s exact test compared anemia rates between the TBV and RBV groups with a 95% confidence interval. Secondary efficacy measures included the SVR defined as HCV RNA <100 copies/mL (39 IU/mL) and/or at least a 2-log decrease from baseline at TW4, TW24, TW48 and FW4 and FW12 and relapse rates at FW4, FW12, and

FW24. Secondary safety measures included the comparison of incidence of treatment-emergent adverse events. Subgroup analysis by HCV RNA levels at baseline, body weight, age, sex, race, and baseline fibrosis were performed using the trend test and the Fisher’s exact test for the primary endpoint. In addition, the Cochran-Mantel-Haenszel KPT-330 research buy procedure, with the

Breslow-Day test was used to examine the homogeneity of treatment effect across strata. The investigators and the sponsor managed the data for this study. The sponsor completed the statistical analysis. The authors had access to the clinical study report and have either written or provided intellectual input to the manuscript. A total of 278 patients were randomized at 51 U.S. centers between March 2007 and October 2008. A total of 86 (41%) of patients in the TBV arms and 25 (36%) in the RBV arm completed treatment and follow-up. Overall, 122 (59%) patients withdrew prematurely

in the TBV arms compared to 45 (64%) in the RBV group. The most commonly cited reasons for premature withdrawal were lack of response (29%) and adverse events (20%). Figure 1 shows the disposition of patients during treatment. Baseline characteristics across the four treatment groups were similar (Table 1). The majority of patients were male (61%) with a mean weight of 82.1 kg and mean age of 49 years. African American or Latino patients accounted for 30% of the study medchemexpress population and 81% had high viral load defined as >400,000 IU/mL at baseline. The proportions of patients in the ITT population with an EVR, the primary endpoint of this study, were comparable between all groups with no statistical difference versus RBV. EVR was achieved in 64.2% (43 of 67) in the 20 mg/kg group, 57.1% (40 of 70) in the 25 mg/kg group, 54.4% (37 of 68) of the 30 mg/kg group and 51.4% (36 of 70) in the RBV group. Virologic response for TW4, 12, 24, and 48 as well as SVR are shown in Table 2. The proportion of patients with undetectable HCV RNA at every time point was similar between the TBV and RBV groups. Although responder rates were numerically lower at TW12 in the TBV 30 mg/kg group and somewhat higher at TW24 and TW48 in the TBV 20 mg/kg group, they were not significantly different for any of the TBV doses compared with RBV.

Materials and Methods: The material comprised 628 final casts for

Materials and Methods: The material comprised 628 final casts for RPDs. Each cast was photographed in a way that would allow the number of existing teeth, the classification of partial edentulism, the RPD support, and the particular parts of the metal framework to be identified. Data collected were analyzed statistically using prevalence tables and the χ2 test. Results: Two hundred seventy six (43.9%) casts were for the maxilla selleck screening library and 352 (56.1%) for the mandible. The most frequently encountered group was Kennedy class I for both arches, while class IV was the classification least encountered (p < 0.001). Of all RPDs constructed,

96.8% had a metal framework (tooth-borne and tooth/tissue-borne), while Y-27632 nmr 3.2% of the RPDs were frameless (tissue-borne, acrylic dentures). The U-shaped palatal connector (horseshoe) in the maxilla

and the lingual bar in the mandible were the most frequently used for all partial edentulism classes, at 55.2% and 95%, respectively. Conclusions: Analysis of the casts revealed that the type of major connectors selected does not comply with the indications for their applications, considering the lack of dental history and clinical examination. This notes the need for further training dentists and dental technicians in aspects of RPD framework design. “
“Cleidocranial dysplasia (CCD) is a rare congenital disorder characterized by skeletal and dental anomalies. This clinical report describes the prosthodontic approach to treating medchemexpress a CCD patient who presented with decreased facial height and relative mandibular protrusion due to maxillary hypoplasia after orthodontic treatment. Functional and esthetic rehabilitation was achieved using telescopic detachable prostheses in the maxilla and osseointegrated implants and metal-ceramic fixed dental prostheses in the mandible. These treatment approaches precluded the need for orthognathic surgical correction and presented a favorable prognosis during the 5-year observation period. Cleidocranial

dysplasia (CCD) is a rare autosomal-dominant skeletal dysplasia best known for its dental and clavicular abnormalities.[1] The most characteristic feature of CCD is hypoplasia or aplasia of the clavicles, resulting in hypermobility of the shoulders.[2] Patients with CCD tend to be of moderately short stature and have proportionally large heads with pronounced frontal and parietal bossing. A broad base of the nose with a depressed nasal bridge as well as ocular hypertelorism may also be observed. The face appears small in relation to the cranium due to the presence of hypoplastic maxillary, lacrimal, nasal, and zygomatic bones.[3] Dental problems are a significant manifestation of CCD. Retained deciduous dentition, delayed eruption or retention of the permanent dentition, and multiple supernumerary teeth are common findings in CCD patients.

Materials and Methods: The material comprised 628 final casts for

Materials and Methods: The material comprised 628 final casts for RPDs. Each cast was photographed in a way that would allow the number of existing teeth, the classification of partial edentulism, the RPD support, and the particular parts of the metal framework to be identified. Data collected were analyzed statistically using prevalence tables and the χ2 test. Results: Two hundred seventy six (43.9%) casts were for the maxilla Selleckchem Panobinostat and 352 (56.1%) for the mandible. The most frequently encountered group was Kennedy class I for both arches, while class IV was the classification least encountered (p < 0.001). Of all RPDs constructed,

96.8% had a metal framework (tooth-borne and tooth/tissue-borne), while VX-809 clinical trial 3.2% of the RPDs were frameless (tissue-borne, acrylic dentures). The U-shaped palatal connector (horseshoe) in the maxilla

and the lingual bar in the mandible were the most frequently used for all partial edentulism classes, at 55.2% and 95%, respectively. Conclusions: Analysis of the casts revealed that the type of major connectors selected does not comply with the indications for their applications, considering the lack of dental history and clinical examination. This notes the need for further training dentists and dental technicians in aspects of RPD framework design. “
“Cleidocranial dysplasia (CCD) is a rare congenital disorder characterized by skeletal and dental anomalies. This clinical report describes the prosthodontic approach to treating MCE a CCD patient who presented with decreased facial height and relative mandibular protrusion due to maxillary hypoplasia after orthodontic treatment. Functional and esthetic rehabilitation was achieved using telescopic detachable prostheses in the maxilla and osseointegrated implants and metal-ceramic fixed dental prostheses in the mandible. These treatment approaches precluded the need for orthognathic surgical correction and presented a favorable prognosis during the 5-year observation period. Cleidocranial

dysplasia (CCD) is a rare autosomal-dominant skeletal dysplasia best known for its dental and clavicular abnormalities.[1] The most characteristic feature of CCD is hypoplasia or aplasia of the clavicles, resulting in hypermobility of the shoulders.[2] Patients with CCD tend to be of moderately short stature and have proportionally large heads with pronounced frontal and parietal bossing. A broad base of the nose with a depressed nasal bridge as well as ocular hypertelorism may also be observed. The face appears small in relation to the cranium due to the presence of hypoplastic maxillary, lacrimal, nasal, and zygomatic bones.[3] Dental problems are a significant manifestation of CCD. Retained deciduous dentition, delayed eruption or retention of the permanent dentition, and multiple supernumerary teeth are common findings in CCD patients.

no NASH Both groups were well matched for clinical parameters (e

no NASH. Both groups were well matched for clinical parameters (e.g. BMI, liver fat, and insulin resistance). Again, no differences were observed in LDL particle size or phenotype or in any lipoprotein subfraction. Conclusion: Patients with NAFLD have a more atherogenic lipo-protein profile (smaller LDL particles and more atherogenic HDL

subparticles) than patients without NAFLD, even when well-matched for BMI and other clinical parameters. We speculate that this lipoprotein profile is driven mostly by buy GSK126 liver fat content and insulin resistance, and appears not to be affected by the severity of liver disease (NASH) or other metabolic features that coexist with NAFLD, such as obesity or hyperglycemia. Disclosures: John Sninsky – Employment: Quest Diagnostics Robert H. Superko – Employment: Celera-Quest Diagnostics Beverly Orsak – Employment: UTHSCSA Kenneth Cusi – Consulting: Merck, Daichi-Sankyo, Roche, Janssen; Grant/ Research Support: Takeda, Novartis, Mannkind The following people have nothing to disclose: Fernando Bril, Arthur M. Baca, Paola Portillo Sanchez, Margaret C. Lo Background and Aim: Although nonalcoholic steatohepati-tis (NASH) is a common liver disease with a risk of progression to cirrhosis, the major cause of morbidity and mortality in NASH is cardiovascular disease (CVD). Oxidative stress and inflammation play a central role in disease progression within the liver but are also independent

predictors of CVD in NASH. Patients with NASH have reduced levels of high density lipoprotein (HDL), which has cardioprotective effects that include HDL dependent reverse cholesterol transport Selleck Pexidartinib (RCT), anti-oxidant and, anti-inflammatory functions. Since insulin resistance, inflammation and oxidative

stress cause HDL dysfunction through alterations of HDL composition, in addition to being medchemexpress key components of NASH pathogenesis, we hypothesized that dyslipidemia-induced hepatic oxidative stress and inflammation in NASH causes loss of RCT and the anti-oxida-tive functions. Methods. HDL functionality was measured using the macrophage cholesterol efflux assay (%) and paraoxonase (PON1) activity (arbitrary units of anti-oxidative function) by a fluorometric assay in apoB-depleted serum from NASH (n=10) and control (n=11) subjects. ATP binding casette subfamily A1 (ABCA1, a protein that transports cholesterol to apoAI forming nascent HDL)-dependent cholesterol efflux was measured in response to induction using 8-Br-cAMP to distinguish ABCA1-dependent and total cholesterol efflux. The ABCA1-de-pendent cholesterol efflux was calculated by substraction of ABCA1-independent from total efflux. Results. NASH patients had higher BMI, HOMA-IR scores, plasma AST, ALT and tri-glycerides while plasma HDLc was non-significanlty lower than controls. However, HDL function quantified by both total and ABCA1-dependent cholesterol efflux capacity of apoB-de-pleted serum from patients with NASH were significantly lower than controls (Total: 15.0±2.4 vs. 19.2.0±2.

2 and 3) Apart from being transcriptionally controlled by NF-κB,

2 and 3). Apart from being transcriptionally controlled by NF-κB, a subset of cytokines/chemokines contains AU-rich elements (AREs) in the 3′NTR of their mRNAs that regulate their expression Selleck JQ1 post-transcritionally

by controlling mRNA stability and/or translation. Among these cytokines/chemokines are IL-6, TNF-α, and MIP-1α,22 and so on, which were found to be induced in HCV-infected 7.5-TLR3 cells (Fig. 1). Although the TLR3-TRIF pathway is less capable of promoting the stabilization of ARE-containing mRNAs than are the myeloid differentiation primary response gene (88)–-dependent pathways,23 further studies are needed to investigate whether ARE-mediated enhancement LY294002 supplier of mRNA stability/translation also contributes to TLR3-dependent up-regulation of specific proinflammatory mediators during HCV infection in hepatocytes. Our study defines the molecular features of HCV PAMP required for TLR3 activation as HCV dsRNA duplexes ≥80-100-bp, regardless of genome location or nucleotide composition. Although it is established that TLR3 senses dsRNA,24 whether TLR3 recognizes HCV RNA duplexes or the secondary structures present

in HCV RNA (such as the stem loops in the core- and NS5B-coding and 3′NTR regions) has not been determined previously. Our data demonstrate that HCV dsRNA duplexes generated during viral RNA replication are the ligands for TLR3 (Fig. 5), whereas the secondary structures present in either +ss or –ss HCV RNAs are not, as evidenced by the finding that neither strand of the NS-3′NTR RNA containing complex stem-loop structures in NS5B and 3′NTR sequences activated RANTES expression (Fig. 5B). Further supporting this, the +ss and –ss HCV RNAs derived from the HCV core-coding region, which contains two stem-loops,17 also failed to activate TLR3, unless the two RNA strands were annealed to form dsRNA duplexes (data not shown). We speculate that the highly structured HCV ssRNAs may not present the perfect

dsRNA structure required for binding to and/or form a stable complex with TLR3, as opposed to HCV dsRNA duplexes. RIG-I preferentially recognizes a segment of HCV MCE公司 3′NTR RNA rich in poly-U/UC nucleotides.11 This is not the case for TLR3, because HCV dsRNA duplexes derived from core, E-p7, NS5A, and NS-3′NTR regions all activated TLR3 with similar potency (Fig. 5C), regardless of their nucleotide compositions. This suggests that the dsRNA duplex structure is the sole determinant for the HCV dsRNA recognition by TLR3, as long as the dsRNA meets the minimal length requirement (see below). We found that HCV dsRNA has to be at least 80-100-bp to reproducibly activate TLR3 and trigger chemokine induction (Fig. 6). This is consistent with the intracellular localization of TLR3 expressed in reconstituted 7.

2 and 3) Apart from being transcriptionally controlled by NF-κB,

2 and 3). Apart from being transcriptionally controlled by NF-κB, a subset of cytokines/chemokines contains AU-rich elements (AREs) in the 3′NTR of their mRNAs that regulate their expression selleck post-transcritionally

by controlling mRNA stability and/or translation. Among these cytokines/chemokines are IL-6, TNF-α, and MIP-1α,22 and so on, which were found to be induced in HCV-infected 7.5-TLR3 cells (Fig. 1). Although the TLR3-TRIF pathway is less capable of promoting the stabilization of ARE-containing mRNAs than are the myeloid differentiation primary response gene (88)–-dependent pathways,23 further studies are needed to investigate whether ARE-mediated enhancement Ipatasertib of mRNA stability/translation also contributes to TLR3-dependent up-regulation of specific proinflammatory mediators during HCV infection in hepatocytes. Our study defines the molecular features of HCV PAMP required for TLR3 activation as HCV dsRNA duplexes ≥80-100-bp, regardless of genome location or nucleotide composition. Although it is established that TLR3 senses dsRNA,24 whether TLR3 recognizes HCV RNA duplexes or the secondary structures present

in HCV RNA (such as the stem loops in the core- and NS5B-coding and 3′NTR regions) has not been determined previously. Our data demonstrate that HCV dsRNA duplexes generated during viral RNA replication are the ligands for TLR3 (Fig. 5), whereas the secondary structures present in either +ss or –ss HCV RNAs are not, as evidenced by the finding that neither strand of the NS-3′NTR RNA containing complex stem-loop structures in NS5B and 3′NTR sequences activated RANTES expression (Fig. 5B). Further supporting this, the +ss and –ss HCV RNAs derived from the HCV core-coding region, which contains two stem-loops,17 also failed to activate TLR3, unless the two RNA strands were annealed to form dsRNA duplexes (data not shown). We speculate that the highly structured HCV ssRNAs may not present the perfect

dsRNA structure required for binding to and/or form a stable complex with TLR3, as opposed to HCV dsRNA duplexes. RIG-I preferentially recognizes a segment of HCV 上海皓元 3′NTR RNA rich in poly-U/UC nucleotides.11 This is not the case for TLR3, because HCV dsRNA duplexes derived from core, E-p7, NS5A, and NS-3′NTR regions all activated TLR3 with similar potency (Fig. 5C), regardless of their nucleotide compositions. This suggests that the dsRNA duplex structure is the sole determinant for the HCV dsRNA recognition by TLR3, as long as the dsRNA meets the minimal length requirement (see below). We found that HCV dsRNA has to be at least 80-100-bp to reproducibly activate TLR3 and trigger chemokine induction (Fig. 6). This is consistent with the intracellular localization of TLR3 expressed in reconstituted 7.